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G Protein-Coupled Receptor Signaling

Methods and Protocols

  • Book
  • © 2019

Overview

Editors:
  1. Mario Tiberi

    1. Ottawa Hospital Research Institute (Neurosciences), Ottawa, Canada

    View editor publications

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  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1947)

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Table of contents (23 protocols)

  1. Front Matter

    Pages i-xv
    Download chapter PDF

  2. Probing Structural Properties of GPCRs

    1. Front Matter

      Pages 1-1
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    2. Assessment of Conformational State Transitions of Class B GPCRs Using Molecular Dynamics

      • Chenyi Liao, Victor May, Jianing Li
      Pages 3-19

    3. Molecular Dynamic Simulations to Probe Water Permeation Pathways of GPCRs

      • Katsufumi Tomobe, Eiji Yamamoto, Kenji Yasuoka
      Pages 21-30

    4. Expression and Purification of a Functional E. coli 13CH3-Methionine-Labeled Thermostable Neurotensin Receptor 1 Variant for Solution NMR Studies

      • Fabian Bumbak, Ross A. D. Bathgate, Daniel J. Scott, Paul R. Gooley
      Pages 31-55

    5. A Combined Cell-Free Protein Synthesis and Fluorescence-Based Approach to Investigate GPCR Binding Properties

      • Anne Zemella, Theresa Richter, Lena Thoring, Stefan Kubick
      Pages 57-77Open Access

  3. Tweaking Ligands for GPCR Studies

    1. Front Matter

      Pages 79-79
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    2. Furan Cross-Linking Technology for Investigating GPCR–Ligand Interactions

      • Marleen Van Troys, Willem Vannecke, Christophe Ampe, Annemieke Madder
      Pages 81-102

    3. Optical Regulation of Class C GPCRs by Photoswitchable Orthogonal Remotely Tethered Ligands

      • Amanda Acosta-Ruiz, Johannes Broichhagen, Joshua Levitz
      Pages 103-136

    4. Chemoselective Acylation of Hydrazinopeptides to Access Fluorescent Probes for Time-Resolved FRET Assays on GPCRs

      • Sridévi M. Ramanoudjame, Lucie Esteoulle, Stéphanie Riché, Jean-François Margathe, Thierry Durroux, Iuliia A. Karpenko et al.
      Pages 137-147

  4. Bioluminescence and Fluorescence Resonance Transfer Energy Approaches

    1. Front Matter

      Pages 149-149
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    2. Time-Resolved FRET-Based Assays to Characterize G Protein-Coupled Receptor Hetero-oligomer Pharmacology

      • Joyce Heuninck, Candide Hounsou, Elodie Dupuis, Eric Trinquet, Bernard Mouillac, Jean-Philippe Pin et al.
      Pages 151-168

    3. Combining Conformational Profiling of GPCRs with CRISPR/Cas9 Gene Editing Approaches

      • Kyla Bourque, Dominic Devost, Asuka Inoue, Terence E. Hébert
      Pages 169-182

    4. Measuring GPCR Stoichiometry Using Types-1, -2, and -3 Bioluminescence Resonance Energy Transfer-Based Assays

      • James H. Felce, John R. James, Simon J. Davis
      Pages 183-197

    5. Combining SRET2 and BiFC to Study GPCR Heteromerization and Protein–Protein Interactions

      • Amina M. Bagher, Melanie E. M. Kelly, Eileen M. Denovan-Wright
      Pages 199-215

    6. Quantification and Comparison of Signals Generated by Different FRET-Based cAMP Reporters

      • Andreas Koschinski, Manuela Zaccolo
      Pages 217-237

  5. Exploring GPCR Signaling Properties

    1. Front Matter

      Pages 239-239
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    2. Measuring GPCR-Induced Activation of Protein Tyrosine Phosphatases (PTP) Using In-Gel and Colorimetric PTP Assays

      • Geneviève Hamel-Côté, Fanny Lapointe, Jana Stankova
      Pages 241-256

    3. Measurement of β-Arrestin Recruitment at GPCRs Using the Tango Assay

      • Geneviève Laroche, Patrick M. Giguère
      Pages 257-267

    4. Probing the Interactome of Corticotropin-Releasing Factor Receptor Heteromers Using Mass Spectrometry

      • Burcu Hasdemir, Juan A. Oses-Prieto, Alma Burlingame, Aditi Bhargava
      Pages 269-285
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Keywords

About this book

This detailed volume assembles comprehensive protocols to assist with the study of structural, molecular, cell biological, and in vivo facets of GPCRs, and to enable the development of experimental tools for screening novel GPCR drugs. Sections explore the tweaking of ligands, bioluminescence and FRET approaches, specific GPCR signaling properties, as well as visualization of subcellular compartmentalization. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and practical, G Protein-Coupled Receptor Signaling: Methods and Protocols serves as an ideal reference for life scientists working in a variety of research fields including molecular pharmacology, cell and developmental biology, brain behavior and physiology, drug development and screening.


Chapter  4 is available open access under a CC BY 4.0 license via link.springer.com.

Editors and Affiliations

  • Ottawa Hospital Research Institute (Neurosciences), Ottawa, Canada

    Mario Tiberi

Bibliographic Information

  • Book Title: G Protein-Coupled Receptor Signaling

  • Book Subtitle: Methods and Protocols

  • Editors: Mario Tiberi

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-9121-1

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2019

  • Hardcover ISBN: 978-1-4939-9120-4Published: 11 April 2019

  • eBook ISBN: 978-1-4939-9121-1Published: 10 April 2019

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XV, 406

  • Number of Illustrations: 16 b/w illustrations, 71 illustrations in colour

  • Topics: Protein Science, Pharmacology/Toxicology, Neurobiology

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